Figure 5. Effect of 5-Aza-dC treatment on ABCB1/Pgp levels and LRPPRC binding to the invMED1 site. (A)ABCB1 mRNA levels were evaluated after 24 h of 0.5 μM 5-Aza-dC treatment. Total RNA was isolated and used in RT-qPCR to determine changes in ABCB1 mRNA levels after normalization to β-actin expression. (B) Representative histograms of Pgp expression after 0.5 μM 5-Aza-dC treatment (1): K562 ctrl and 5-Aza-dC-treated K562 cells (2): Lucena ctrl and Lucena cells treated with 5-Aza-dC. PE-isotype antibody was used as a control. (C) RT-qPCR quantification of LRPPRC binding to the ABCB1 promoter in 5-Aza-dC-treated K562 and Lucena cells. DNA amplification was quantified in bound and unbound fractions after normalization with SMAD8 nonspecific amplification. Normalized fractions were used to calculate the bound/input ratio. The results are expressed as the mean ± SD for three independent experiments. Ctrl: control; K5: K562; LU: Lucena.